Mitosis : methods and protocols / edited by Andrew D. McAinsh. — Dordrecht ; London : Humana Press, 2009. – (58.17/M592/v.545)

Contents

    Contents
    
    Preface
    Contributors ix
    1 Knock-in and Knock-out: The Use of Reverse Genetics in Somatic Cells to Dissect Mitotic Pathways 1
    2 Functional Dissection of Mitotic Regulators Through Gene Targeting in Human Somatic Cells 21
    3 RNAi in Drosophila $2 Cells as a Tool for Studying Cell Cycle Progression 39
    4 Production of Mitotic Regulators Using an Autoselection System for Protein Expression in Budding Yeast 63
    5 Hydrodynamic Analysis of Human Kinetochore Complexes During Mitosis 81
    6 Isolation of Protein Complexes Involved in Mitosis and Cytokinesis from Drosophila Cultured Cells 99
    7 Automated Live Microscopy to Study Mitotic Gene Function in Fluorescent Reporter Cell Lines 113
    8 Electron Tomography of Microtubule End-Morphologies in C. elegans Embryos 135
    9 Dissecting Mitosis with Laser Microsurgery and RNAi in Drosophila Cells 145
    10 Fluorescence Imaging of the Centrosome Cycle in Mammalian Cells 165
    11 Visualization of Fluorescence-Tagged Proteins in Fission Yeast: The Analysis of Mitotic Spindle Dynamics Using GFP-Tubulin Under the Native Promoter 185
    12 Analysing Kinetochore Function in Human Cells: Spindle Checkpoint and Chromosome Congression 205
    13 Probing Kinetochore Structure and Function Using Xenopus laevis Frog Egg Extracts 221
    14 Live Cell Imaging of Kinetochore Capture by Microtubules in Budding Yeast 233
    15 The Spindle Checkpoint: Assays for the Analysis of Spindle Checkpoint Arrest and Recovery 243
    16 Measuring Proteolysis in Mitosis 259
    17 An In Vitro Assay for Cdc20-Dependent Mitotic Anaphase-Promoting Complex Activity from Budding Yeast 271
    18 In Vitro Assays for the Anaphase-Promoting Complex/Cyclosome (APC/C) in Xenopus Egg Extracts 287
    19 Preparation of Synchronized Human Cell Extracts to Study Ubiquitination and Degradation 301
    20 Biochemical Analysis of the Anaphase Promoting Complex: Activities of E2 Enzymes and Substrate Competitive (Pseudosubstrate) Inhibitors 313
    Index 331